fibroblasts on beads

Daniel Lottaz wrote:
>> I intend to culture fibroblasts on beads prior to FACS-Analysis. Anyone
> experience in such a technique?
>> Many thanks for any contribution.
>> Daniel Lottaz, Berne
> e-mail: daniel.lottaz at zmk.unibe.ch
Fibroblasts should not be difficult to culture on microcarrier
beads. I routinely use Pharmacia Cytodex Type III (collagen type 1
coated) mc-beads to culture McCoy cells for subsequent infection with
Chlamydia trachomtis. There are two texts that come to mind for general
information, the first being R. Ian Freshney "Culture of Animal Cells"
Wiley Liss ISBN 0-471-58966-7, the second being a technical manual
"Microcarrier cell culture" published by Pharmacia.
I have also cultured WI-38 and primary human uterine stroma on
the Cytodex -3 beads with very good results.
There is a certain amount of fine tuning required for obtaining
the best culture conditions but it's pretty straight-forward. One
pitfall is that there because there is usually a higher density of cells
for the volume of medium in the flask, the pH drop and rate of medium
depletion is faster than stationary cultures.
I hope this helps and the mc bead approach works for you.
Stephen Knight